中药材蟾衣基原的分子鉴定

Molecular Identification of a Traditional Chinese Medicine Toad Slough

为了探索建立快速实用的蟾衣基原分子鉴定方法,本研究以新鲜蟾衣为材料,显微观察浸润后蟾衣的外部形态特征,并以两栖纲蟾蜍科动物线粒体基因组DNA的细胞色素氧化酶Ⅰ基因的内部间隔序列两端的保守序列设计引物,应用DNA条形码技术对蟾衣的基原动物进行分子鉴定。经形态学观察与分子鉴定,本研究蟾衣药材的外部形态与中华大蟾蜍相似,其mtCOⅠ条形码序列与GenBank收录的中华大蟾蜍序列相似度在99%以上,聚类分析结果显示其和中华大蟾蜍聚在一个分支上。本研究首次应用分子鉴定的方法对蟾衣进行鉴定,建立了实用有效的蟾衣分子鉴定的方法,突破了蟾衣鉴定的难关,保证了中药蟾衣在中医临床上的安全用药,为名贵中药材的鉴定提供理论依据。

In order to establish a rapid and practical method for the molecular identification of the basidiomata of toad coats,this study was carried out by microscopically observing the external morphological characteristics of toad coats after infiltration with fresh toad skin and designing primers based on the conserved sequences of the internal spacer of the cytochrome oxidaseⅠgene of the mitochondrial genomic DNA of the amphibian Toadidae,and applying the DNA barcoding technique for the molecular identification of the basidiomata of toad coats.The mtCOⅠbarcode sequences were more than 99%similar to those of the Chinese toad in GenBank,and the clustering analysis showed that they were clustered on the same branch as the Chinese toad.In this study,the molecular identification method was applied for the first time to identify the toadstool,which is a breakthrough in the identification of the toadstool,ensuring the safe use of the toadstool in traditional Chinese medicine clinics and laying the foundation for the identification of valuable Chinese herbs.